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Microtransferrinuria and microalbuminuria: enhanced immunoassay.
C P McCormick1, Z K Shihabi, J C Konen
1Department of Pathology, Wake Forest University, Winston-Salem, NC 27103.
Annals of Clinical and Laboratory Science
|November 1, 1989
Summary
A new method enhances urine protein detection by concentrating samples with trichloroacetic acid. This improved immunoassay accurately measures urinary microtransferrin and microalbumin, aiding in disease diagnosis.
Area of Science:
- Clinical Chemistry
- Biochemistry
- Analytical Chemistry
Background:
- Accurate quantification of urinary proteins like microtransferrin and microalbumin is crucial for diagnosing kidney diseases.
- Existing immunoassay methods may lack sensitivity or require complex sample preparation.
Purpose of the Study:
- To develop and validate an enhanced-sensitivity immunoassay for urinary microtransferrin and microalbumin.
- To establish a reliable method for concentrating urine samples while removing interfering substances.
Main Methods:
- Protein precipitation using cold trichloroacetic acid (TCA) followed by redissolution in phosphate buffer.
- 10-fold sample concentration and chromogen removal.
- Assay of concentrated samples using immunoturbidity and radial immunodiffusion.
Main Results:
- The TCA precipitation method achieved an average recovery of 82% for microtransferrin and 91% for microalbumin.
- Established reference ranges for normal adults: 0-0.9 mg/g creatinine for microtransferrinuria and 5-32 mg/g creatinine for microalbuminuria.
- Demonstrated applicability to other urinary or cerebrospinal fluid proteins like B2-microglobulin.
Conclusions:
- The developed TCA-based precipitation method significantly enhances the sensitivity of urinary microtransferrin and microalbumin immunoassays.
- This method offers a robust and efficient approach for concentrating protein samples, improving diagnostic accuracy.
- The technique is versatile and can be adapted for the analysis of other protein biomarkers.